Characterization of pathogenic factors for premenstrual dysphoric disorder using machine learning algorithms in rats.

We established a methodology using machine learning algorithms for determining the pathogenic factors for premenstrual dysphoric disorder (PMDD). PMDD is a disease characterized by emotional and physical symptoms that occurs before menstruation in women of childbearing age. Owing to the diverse manifestations and various pathogenic factors associated with this disease, the diagnosis of PMDD is time-consuming and challenging. In the present study, we aimed to establish a methodology for diagnosing PMDD. Using an unsupervised machine-learning algorithm, we divided pseudopregnant rats into three clusters (C1 to C3), depending on the level of anxiety- and depression-like behaviors. From the results of RNA-seq and subsequent qPCR of the hippocampus in each cluster, we identified 17 key genes for building a PMDD diagnostic model using our original two-step feature selection with supervised machine learning. By inputting the expression levels of these 17 genes into the machine learning classifier, the PMDD symptoms of another group of rats were successfully classified as C1-C3 with an accuracy of 96%, corresponding to the classification by behavior. The present methodology would be applicable for the clinical diagnosis of PMDD using blood samples instead of samples from the hippocampus in the future.

Bidirectional effects of voluntary exercise on the expression of Bdnf isoforms in the hippocampus of Hatano rat strains displaying different activity levels.

Brain-derived neurotrophic factor has functional mRNA isoforms, whose expression is assumed to mediate the beneficial effects of exercise in neuropsychiatric disorders. This study aims to reveal the mechanism of intensity-dependent effects of voluntary exercise, focusing on the expression of Bdnf mRNA isoforms in Hatano rats. Animals with different voluntary activity were housed in cages with a locked or unlocked wheel for 5 weeks. The expression levels of Bdnf isoforms and the corresponding coding sequences (CDS) were measured in the hippocampus using real-time polymerase chain reaction (PCR). We found that exercise increased the expression of Bdnf isoform containing exon 1 in the high-intensity-running strain and decreased the expressions of Bdnf exon 1, 3, 6, 7, 8, and 9a in mild-intensity-running animal. The expression of Bdnf CDS was increased by exercise in both strains. These results suggest that expressions of Bdnf isoforms depend on the intensities of voluntary exercise, but the involvement of subjects' genetic background could not be excluded. Our finding also implies that the bidirectional effects of exercise may not be mediated via the final product of Bdnf.

Possible effects of voluntary exercise intensity on anxiety-like behavior and its underlying molecular mechanisms in the hippocampus: Results from a study in Hatano rats.

The prevalence of neuropsychiatric diseases, including anxiety disorders, has increased in recent years. A better understanding of the mechanisms mediating symptoms in these disorders is essential for developing treatments. Although voluntary exercise can alleviate symptoms, its anxiolytic effect varies with the intensity of the activity. Therefore, to investigate the usefulness of voluntary exercise in alleviating the symptoms of neuropsychiatric disorders, assessing its effect based on intensity is required. Hatano rats, consisting of high- and low-avoidance animals (HAA and LAA, respectively), differ in their propensity to voluntary exercise. These animals are useful for examining the effects of voluntary running activity differing in intensity on anxiety-like behavior. We housed Hatano rats in cages containing locked or unlocked running wheels starting at 4 weeks of age, conducted elevated plus maze test at 8 weeks of age, followed by plasma corticosterone measurement and DNA microarray analysis on hippocampal tissue at 9 weeks of age. Our results show that only LAA (mild-intensity running animals), but not HAA (high-intensity running animals), had reduced anxiety-like behavior without plasma corticosterone change. In addition, LAA had increased immunity-related gene expression, but decreased proteolysis-related gene expression. Our findings suggest that mild-intensity voluntary running mediates the anxiolytic effect of exercise and is regulated through increasing the expression of immunity-related genes or decreasing the expression of proteolysis-related genes in the hippocampus.

Toxicological effects of Tris (1,3-dichloro-2-propyl) phosphate exposure in adult male rats differ depending on the history of exposure in the neonatal period.

There is increasing concern about multiple high concentration exposure to toxins in disaster and emergency situations. However, conventional toxicology testing methods may not adequately address these situations. Thus, we assessed whether the toxic effects of exposure in the adulthood differ depending on the presence or absence of neonatal exposure to Tris (1,3-dichloro-2-propyl) phosphate (TDCIPP) in male rats to investigate the effects of exposure history of chemicals. In the neonatal stage [postnatal days (PNDs) 1-7], animals were treated with either sesame oil (5 ml/kg/day) as a control or TDCIPP (250 mg/kg/day) dissolved in sesame oil. In adulthood (PND 101-107), animals were treated with either sesame oil (5 ml/kg/day) or TDCIPP (650 mg/kg/day). One day after the final administration, dissection was performed, and body and organ weight, hematology, blood biochemistry, and histopathology were examined. The results demonstrated that the toxic effects of TDCIPP exposure in adulthood on adrenal gland size, serum iron content, and unsaturated iron binding capacity were enhanced by TDCIPP exposure in the neonatal stage. From these findings, it was indicated that the toxic effects of TDCIPP exposure in the adult stage are affected by pediatric exposure. These results suggest that the toxic effects of high-dose and long-term unsteady exposure to chemicals in large-scale disasters may change based on the exposure history of chemicals.

Effects of tris(1,3-dichloro-2-propyl) phosphate on epididymal sperm parameters in adult male rats.

Tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) is widely used as a flame retardant and is known to exhibit anti-androgenic effects in vitro and in vivo. To assess the reproductive toxicity potency of TDCIPP, we investigated the effects of 7 days of TDCIPP oral administration on epididymal sperm motion and concentration in adult male Wistar-Imamichi rats. Thirty-five days after the final administration, sperm parameters were evaluated by computer-assisted sperm analysis. Results showed that sperm swimming progression and vigor and sperm concentration in TDCIPP-treated rats were unexpectedly higher than those in control rats. TDCIPP did not significantly affect the percentage of motile sperms or sperm swimming pattern. These results contribute to the understanding of the biological effects of TDCIPP.

Novel toxicity of tris(1,3-dichloro-2-propyl) phosphate in adult male rats.

The widespread use of tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) as a flame retardant has led to its release to the environment. Thus, the toxicological effects of TDCIPP on humans and animals are of importance. For better understanding of its potential toxicities, TDCIPP (250, 500, or 650 mg/kg/day) or vehicle control was administrated orally to adult male Wistar-Imamichi rats for 7 days. After the final administration of compounds, organ weights, histopathology, blood biochemistry, and hematology were examined. Hepatic toxicity was observed at doses ≥ 500 mg/kg/day of TDCIPP, and renal toxicity was observed at 650 mg/kg/day. The anti-androgenic activity of TDCIPP was previously confirmed in vitro and in vivo, but weights of epididymis, an androgen-dependent organ, were not affected by TDCIPP treatment in adults. Serum alkaline phosphatase activity was significantly decreased in all TDCIPP-treated rats independent of dose. Hemoglobin concentration, hematocrit, red blood cell count, and reticulocyte count were decreased in all TDCIPP-treated rats, but mean corpuscular volume, total iron-binding capacity, and serum iron were normal, suggesting that renal anemia was caused by TDCIPP. Together with previous reports on effects of anti-androgenic substances on red blood cell indices, anemia caused by TDCIPP could be due to its anti-androgenic activity. These considerations will contribute to further assessment of the toxicity of the compound.

Temporal order memory of the rat in spontaneous object recognition: effects of number of items, exposure interval, and retention time.

Temporal order memory was analyzed using a spontaneous object recognition (SOR) paradigm in rats. In SOR, animals were allowed to explore freely two or five different objects sequentially in the sample phase, and then, two different objects shown in the sample phase were simultaneously presented, and exploration time of object shown earlier compared with that shown later was regarded as a discrimination index. Here we investigated the effects of (1) number of sample items, (2) sample exposure interval, and (3) retention time between the sample and test phases on temporal order memory in SOR. In experiment 1, rats showed significant discrimination between the objects presented earlier and later under five-item, but not two-item, condition. In experiment 2, using the five-item condition, longer retention time (between sample and test phases) induced lower discrimination and longer sample exposure interval produced higher discrimination. In experiment 3, using the two-item condition, longer sample exposure interval produced higher discrimination and longer retention time tended to make rats' discrimination worse. Taken together, these results indicate that rats' performance in this test paradigm was better with more sample items and longer exposure intervals as well as shorter retention time, suggesting that the familiarity to items and the temporal distance from the present to the occurrence of events affect temporal order memory.